This proposal consists of three projects: I - Examination of mechanisms which govern the tissue specific and coregulated expression of the alpha1(V), alpha2(V), and alpha1(III) collagen genes. Experimental approaches include 1) sequence determination for the 5' portion of the human alpha1(V) gene and 2) functional assays of 5' flanking and first intron sequences of the three genes in cultured cells known to express the alpha1(V) and alpha2(V) genes in a tissue-specific fashion, followed by binding assays in nuclear extracts of the same cells. II - Investigation of developmental and structural roles for the alpha1(V) gene by: a) identification of polymorphisms in the alpha1(V) gene which cosegregate with nail-patella syndrome and/or tuberous sclerosis, two heritable diseases which map to the same chromosomal location as the alpha1(V) gene; b) characterization of mutations in the alpha1(V) gene of patients; and c) isolation of a cosmid containing the gene for study of intron/exon organization, and for mutagenesis and expression in transgenic mice. III - Study of a collagen chain, nominally alpha1(XVI), discovered in my lab by cDNA cloning, which is coexpressed with the alpha1(V) and alpha5(IV) collagen chains in a line of lung cells. This chain has some, but lacks other features common to known type IV chains. Thus, it is either representative of a previously unknown collagen type, or is a new type IV collagen chain. Goals include determining the complete primary structure and tissue distribution of this new chain, and examining the possibility that it is complexed to the alpha5(IV) chain. Standard molecular biological, immunochemical, and biochemical methods will be employed. Chromosome mapping of the human alpha1(XVI) gene will also be performed. These studies will increase our understanding of both new and relatively uncharacterized collagen types, which are likely to play a variety of roles in normal cell/ECM interactions and to be involved in certain diseases, hereditary or acquired.